QIAGEN Sequencing Services — Single-Read ServicesSingle-read sequencing services (96-well format) are ideal for small-scale analysis of cDNAs and ESTs, identification of clones, and verification of vector/insert transitions. All single read services include extensive documentation of the sequencing data, which is available via e-mail, FTP file, or on CD.

Easy-read sequencing Easy-read sequencing is the service of choice for standard sequencing reactions. This highly automated service allows rapid processing of purified DNA samples. Easy-read sequencing is offered as tube service, or as a plate service for sequencing of samples in batches of 96. Easy-read sequencing does not include template purification (i.e., purification of PCR fragments or plasmid preparations prior to sequencing reaction setup). Sequencing reactions are set up using the DNA concentration information provided by the customer. All sequencing results are subjected to quality control using Phred20 algorithms (typically 600–900 bases of Phred20 controlled sequence are obtained). Quality documentation including raw data and quality clipped sequence data is delivered via e-mail, ftp, or on CD. Failed sequences are not repeated. This service provides high-quality single-stranded sequencing data 1–2 working days after receipt of purified DNA.

Single-read sequencing
If your sequencing reaction needs special attention we recommend our single-read sequencing Service. The sequencing results are manually checked and any failed sequencing reactions are repeated using modified sequencing parameters. This means that you get the most reliable service even from difficult samples (e.g., samples with high GC content). This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. Single-read sequencing with template preparation includes the purification of PCR fragments or plasmids prior the sequencing reaction setup. Templates are quality controlled to ensure an optimal sequencing reaction setup. All sequencing results are subjected to quality control using Phred20 algorithms (typically 600–900 bases of Phred20 controlled sequence are obtained). Quality documentation including raw data and quality clipped sequence data is delivered via e-mail, ftp, or on CD. Failed sequences are inspected by our experienced team and repeated using modified sequencing parameters. This service provides high-quality single-stranded sequencing data in 2–3 working days (3–4 working days with template purification).

BAC end-sequencing End-sequencing of BACs, PACs, or cosmids. The results are manually checked by our experienced team and failed sequencing reactions are repeated using modified sequencing parameters. This means that you get the most reliable service even from difficult samples (e.g., samples with high GC content). This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. The delivery time is approximately 2–3 working days (3–4 working days with template purification).

siRNA hairpin run
siRNA expression vector templates exhibit secondary structures that are often difficult to sequence. QIAGEN has developed a special sequencing reaction that enables accurate sequencing of siRNA expression vector templates. The results are manually checked by our experienced team and failed sequencing reactions are repeated using modified sequencing parameters. This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. The delivery time is approximately 2–3 working days (3–4 working days with template purification).

Verification sequencing Verification sequencing provides confirmation of an already known DNA sequence, as well as detection of mutations or deletions. Both DNA strands are completely sequenced using custom primers based on a provided DNA sequence. The sequence is verified against the provided reference sequence.

Verification sequencing provides:

Template purification

Design and synthesis of internal primers

Complete sequencing of both DNA strands (>99.995% accuracy)

Sequence editing and assembly

Verification of the sequence against a customer-provided reference sequence

Sequence data in publication-ready format

Project documentation

Electronic data delivery

Experienced and reliable support

Alternatively, BACs, PACs, or cosmids can be sequenced to a predetermined sequence coverage including assembly. Please ask for this service.

Verification sequencing Under GLP-like conditions
Verification sequencing Under GLP-like conditions provides confirmation of an already-known DNA sequence including comprehensive documentation of the sequencing results. Both DNA strands are completely sequenced to at least 4-fold coverage. The sequence is verified against a provided reference sequence. Documentation can be used for patent applications, litigations, or FDA submissions.

GLP-like verification sequencing comprises:

Template purification

Design and synthesis of internal primers

Complete sequencing of both DNA strands (>99.995% accuracy)

4-fold coverage

Sequence editing and assembly

Verification of the sequence against a customer-provided reference sequence

De novo publication-ready sequencing The de novo publication-ready sequencing service provides publication-ready DNA sequence from unknown template DNA. Both DNA strands are completely sequenced and the sequence is fully edited and assembled. Depending on the project size, different sequencing strategies can be applied (i.e., primer walking for plasmids or shotgun sequencing cosmids, BACs, or PACs).

If you provide us with an E. coli host strain harbouring the construct of interest, we will apply the best strategy to determine the sequence and we will send back the sequencing results within 20 working days.

De novo publication-ready sequencing provides:

Complete sequencing of both DNA strands (>99.995% accuracy)

Template purification

Design and synthesis of internal primers or shotgun library generation

Sequence editing and assembly

Sequence data in a publication-ready format

Applicable project documentation

Electronic data delivery

Experienced and reliable support

Alternatively, BACs, PACs, or cosmids can be sequenced to a predetermined sequence coverage including assembly. Please ask for this service.

De novo investigation-grade sequencing
De novo investigation-grade sequencing provides a single stranded primer walking approach for preliminary analysis of unknown sequences. The target DNA is sequenced to single-strand coverage and the sequence is edited and assembled. We guarantee a sequence quality score of Phred20 (i.e., an error rate of 1:100 nucleotides) for all assigned bases.

De novo investigation-grade sequencing provides:

Complete sequencing of one DNA strand (>99% accuracy)

Template purification

Design and synthesis of internal primers

Sequence editing and assembly

Sequence data in a publication-ready format

Applicable project documentation

Electronic data delivery

Experienced and reliable support

De novo publication sequencing under GLP-like conditions

De novo publication sequencing under GLP-like conditions provides double stranded sequencing including comprehensive documentation. Both DNA strands are completely sequenced to at least four fold coverage. Documentation can be used for patent applications, litigations, or FDA submissions.

If you provide us with an E. coli host strain harbouring the construct of interest, we will apply the best strategy to determine the sequence and we will send back the sequencing results within 20 working days.

De novo publication sequencing Under GLP-like conditions provides:

Complete sequencing of both DNA strands (>99.995% accuracy)

At least 4-fold sequencing coverage

Template purification

Design and synthesis of internal primers or shotgun library generation

QIAGEN has participated in various genome projects. Our genome-sequencing service includes the production of BAC or shotgun-cloned libraries or whole genome shotgun libraries, high-throughput sequencing, and data annotation.

QIAGEN sequencing services has become a well-established participant in many publicly funded and private genome projects. The following table shows our contribution to selected projects:

One critical factor for successful genome sequencing is the generation of whole genome shotgun libraries. Our shotgun libraries are of an extremely high quality. Shotgun libraries with different insert sizes are used to establish the sequence either up to publication quality (>99.995% sequence accuracy) or up to a certain genomic coverage.

We also offer various bioinformatic analyses including complete annotation and comparative sequencing.

For large-scale projects, a dedicated project management plan and team is instigated to carry out all necessary work in due speed and to the highest accuracy and quality levels.

QIAGEN Sequencing Services — Special Services

QIAGEN offers a number of special services that complement our sequencing service. These services include the sequencing of templates with a high degree of secondary structure, the sequencing of genomic DNA, the detection of mutations, DNA purification, whole genome amplification, and bioinformatic analysis.

siRNA hairpin run siRNA expression vector templates exhibit secondary structures that are often difficult to sequence. QIAGEN has developed a special sequencing reaction that enables accurate sequencing of siRNA expression vector templates. The results are manually checked by our experienced team and any failed sequencing reactions are repeated using modified sequencing parameters. This service is offered with or without template purification as a plate service for sequencing of samples in batches of 96. The delivery time is approximately 2–3 working days (3–4 working days with template purification).

DNA purification service Don't waste your time with routine work. QIAGEN offers both small- and large-scale DNA preparation services from a variety of different starting materials.

The DNA purification service provides:

Plasmid DNA preparation from bacterial cultures

Cosmid, fosmid, BAC preps

PCR purification

DNA purification from EDTA-blood

DNA purification from buccal swabs

Preparation in 96- or 384-well plate format

Whole genome amplification service If you do not have enough starting material for your downstream analysis, we recommend amplification of your genomic DNA. The REPLI-g service — based on proven REPLI-g whole genome amplification technology — allows the amplification of unlimited amounts of DNA from limited samples with minimal sequence bias. A stringent quality control assay provides information on the quality of the amplified DNA, enabling reliable predictions for the success of your downstream assay to be made. For more information, or to find out how to take advantage of this service click here.

Bioinformatic analysis Every sequencing project has special requirements for bioinformatic resources (e.g., large scale BLAST analysis, EST clustering, custom primer design, or annotation). The QIAGEN Sequencing Service provides in-depth bioinformatic analysis of DNA sequences for sequencing projects of any scale

Principle

Simply send your sample and take advantage of QIAGEN's expertise. From sample receipt, DNA purification and QC of purified DNA to sequencing and data analysis and delivery, QIAGEN offers a one-stop service for all your contract manufacturing and sequencing needs.

Procedure

To get started, obtain a quotation for your project request. If you are a new customer, please contact QIAGEN Sequencing Services (QSS) for pricing information.

Complete an order request Please download our order form in Excel format. Order forms in excel format should be e-mailed to sequencing@qiagen.com in advance. A hardcopy MUST BE submitted with every package shipped to QIAGEN.

Sample submission and shipping Please provide the following DNA concentrations per sequencing reaction:

Plasmid (3-10 kb): 0.5 µg

Plasmid (10-20 kb): 1 µg

Cosmid (30-45 kb): 3 µg

BAC/ PAC: 3 µg

Bacterial genomic DNA: 10 µg

PCR product (< 500 bp): 0.1 µg

PCR product (500-2000 bp): 0.2 µg

PCR product (>2000 bp): 0.5 µg

Unpurified PCR product: send the whole PCR reaction (50-100 µl)

If you submit your own primers, we need 20 pmol primer per sequencing reaction. Please supply your primer dissolved in water (5 pmol/µl),

Procedures for shipping clones and DNA from countries outside the EU to QIAGEN Sequencing Services
QIAGEN has selected FedEx as our preferred carrier to Germany.

Please follow this checklist carefully to ensure that your samples are processed efficiently through FedEx and German customs.

A padded shipping envelope may be used for small numbers of tubes or plates. For larger shipments, boxes may be used but must accommodate the FedEx International Waybill and Commercial Invoice.

Securely pack samples for shipment with padding to prevent damage to vials or culture plates. Tighten caps on liquid samples and seal with Parafilm to prevent leakage. No samples with a biohazard rating above the lowest level (i.e., BL1, S1, P1, L1) should be shipped. Clones from non-infectious organisms in standard laboratory strains of E. coli are all acceptable.

On a FedEx International Air Waybill, write in the date of shipment, your account number, and your complete address and phone number in box 1.

In box 2, enter: André Bahr

Phone +-49-2103-29-16345

QIAGEN Genomic Services / Sequencing

QIAGEN GmbH

Max-Volmer-Str. 4

Hilden. Germany D-40724

Enter a description and harmonized code for the contents of your shipment in box 3. For bacterial cultures, enter “3002.90.50 Microorganisms, E. coli cultures, non-infectious, non-hazardous, for research purposes only, contained in (give number) vials/plastic plates.” For DNA samples (e.g. genomic DNA or PCR products), write “2933.39.90 Nucleic Acids, contained in (give number) vials/plastic plates.” The value of the shipment stated for customs should not total more than $25 per shipment. (This is basically the value of the plastics; the clones themselves, although of course worth a lot to you, are considered by customs as having no value. The $25 or less value makes it easier to get the shipment through customs, as there will be no duties applied.) The box marked “No SED required, value $2500 or less” should be checked.

In box 4, select the shipping option that works best for you. Please use FedEx First or FedEx Priority for shipments containing dry or wet ice.

In box 5, indicate your packaging option. No special handling should be indicated in box 6 if package is shipped early in the week. We recommend that all shipments containing dry or wet ice be shipped on either Monday or Tuesday.

On a Commercial Invoice form, enter the information requested to match the information on the FedEx International Waybill. In particular, enter the exact same description used in box 4 under "Description of Goods". Under “No. Units” write the number of plates or vials, under "Unit Value", write the value of each plate or vial, and provide the multiplied value in the total value column.

Under “Terms of Sale (Incoterm)”, write “CIF”. This indicates that the basis of the “sale” (any international transfer) is cost, insurance, and freight. Fill in the number of packages, weight, shipping cost, and insurance ($0).

Sign your name under "Shipper’s Signature".

Insert the International Waybill and two copies of the Commercial Invoice in a plastic adhesive FedEx envelope and affix it to your shipping envelope or other container. Call FedEx Customer Service (1-800-Go-FedEx) for a pick up or your nearest drop off location. For further shipping information, contact FedEx International Services (800-247-4747).

IMPORTANT: Do not forget to indicate the sender when you send your samples! This information is essential so that we can correctly match your samples to your order form. Alternatively, please include a hard copy of the order form when you send your samples to us.
You will receive an acknowledgement of your order by e-mail when we receive your samples.
If you have any questions, please do not hesitate to call your local Technical Service or the Genomic Services Department in Germany at +49 2103 29 16234 or +49 2103 29 16345.